Benzylether and benzylamino beta-secretase inhibitors for the treatment of alzheimer&#39;s disease

ABSTRACT

The present invention is directed to compounds of formula (I) 
                         
which are inhibitors of the beta-secretase enzyme and that are useful in the treatment of diseases in which the beta-secretase enzyme is involved, such as Alzheimer&#39;s disease. The invention is also directed to pharmaceutical compositions comprising these compounds and the use of these compounds and compositions in the treatment of such diseases in which the beta-secretase enzyme is involved.

CROSS-REFERENCE TO RELATED APPLICATIONS

This application claims priority under 35 U.S.C. §119(e) fromProvisional Application Ser. No. 60/508,369, filed Oct. 3, 2003.

REFERENCE TO JOINT RESEARCH AGREEMENT

This invention was made as a result of activities undertaken within thescope of a Joint Research Agreement between Merck & Co., Inc. andSunesis Pharmaceuticals, Inc.

BACKGROUND OF THE INVENTION

Alzheimer's disease is characterized by the abnormal deposition ofamyloid in the brain in the form of extra-cellular plaques andintra-cellular neurofibrillary tangles. The rate of amyloid accumulationis a combination of the rates of formation, aggregation and egress fromthe brain. It is generally accepted that the main constituent of amyloidplaques is the 4 kD amyloid protein (βA4, also referred to as Aβ,β-protein and βAP) which is a proteolytic product of a precursor proteinof much larger size. The amyloid precursor protein (APP or AβPP) has areceptor-like structure with a large ectodomain, a membrane spanningregion and a short cytoplasmic tail. The Aβ domain encompasses parts ofboth extra-cellular and transmembrane domains of APP, thus its releaseimplies the existence of two distinct proteolytic events to generate itsNH₂- and COOH-termini. At least two secretory mechanisms exist whichrelease APP from the membrane and generate soluble, COOH-truncated formsof APP (APP_(S)). Proteases that release APP and its fragments from themembrane are termed “secretases.” Most APP_(S) is released by a putativex-secretase which cleaves within the Aβ protein to release α-APP_(S) andprecludes the release of intact Aβ. A minor portion of APP_(S) isreleased by a β-secretase (“β-secretase”), which cleaves near theNH₂-terminus of APP and produces COOH-terminal fragments (CTFs) whichcontain the whole Aβdomain.

Thus, the activity of β-secretase or β-site amyloid precursorprotein-cleaving enzyme (“BACE”) leads to the abnormal cleavage of APP,production Aβ, and accumulation of β amyloid plaques in the brain, whichis characteristic of Alzheimer's disease (see R. N. Rosenberg, Arch.Neurol., vol. 59, September 2002, pp. 1367-1368; H. Fukumoto et al,Arch. Neurol., vol. 59, September 2002, pp. 1381-1389; J. T. Huse et al,J. Biol. Chem., vol 277, No. 18, issue of May 3, 2002, pp. 16278-16284;K. C. Chen and W. J. Howe, Biochem. Biophys. Res. Comun, vol. 292, pp702-708, 2002). Therefore, therapeutic agents that can inhibitβ-secretase or BACE may be useful for the treatment of Alzheimer'sdisease.

The compounds of the present invention are useful for treatingAlzheimer's disease by inhibiting the activity of β-secretase or BACE,thus preventing the formation of insoluble Aβ and arresting theproduction of Aβ.

SUMMARY OF THE INVENTION

The present invention is directed to compounds that are inhibitors ofthe β-secretase enzyme, and that are useful in the treatment of diseasesin which the β-secretase enzyme is involved, such as Alzheimer'sdisease. The invention is also directed to pharmaceutical compositionscomprising these compounds and the use of these compounds andcompositions in the treatment of such diseases in which the β-secretaseenzyme is involved.

DETAILED DESCRIPTION OF THE INVENTION

The present invention is directed to compounds of the formula (I):

wherein:

-   X is O or NH;-   Y is CH or N;-   R¹ is (1) aryl selected from the group consisting of phenyl and    napthyl, or (2) heterocyclyl selected from the group consisting of    piperazinyl, piperidinyl, pyrrolidinyl, pyrazinyl, dihydropyrazinyl,    pyrazolyl, dihydropyrazolyl, pyridazinyl, pyridyl, dihydropyridinyl,    pyrimidinyl, dihydropyrimidinyl, pyrrolyl, dihydropyrrolyl,    tetrazolyl, dihydrotetrazolyl, furanyl, dihydrofuranyl,    tetrahydrofuranyl, imidazolyl, dihydroimidazolyl, triazinyl,    pyranyl, tetrahydropyranyl, thiazolyl, thienyl, dihydrothienyl,    thiophenyl, triazolyl, dihydrotriazolyl, morpholinyl,    thiomorpholinyl, dihydrothiadiazolyl, tetrahydrothienyl, oxazolyl,    isoxazolyl, thiazolyl, oxadiazolyl, indolyl, quinolinyl,    isoquinolinyl, benzimidazolyl and benzoxazolyl,    -   wherein said aryl or heterocyclyl is unsubstituted or        substituted with one or more        -   (a) halo,        -   (b) —C₁₋₆alkyl,        -   (c) —C₂₋₆ alkenyl,        -   (d) —C₂₋₆ alkynyl,        -   (e) —OH,        -   (f) —CN, or        -   (g) —O—C₁₋₆alkyl;-   R² is selected from the group consisting of:    -   (1) R⁴—S(O)₂N(R⁷)—, wherein R⁴ is C₁₋₆alkyl, wherein said alkyl        is unsubstituted or substituted with one or more        -   (a) halo,        -   (b) —C₁₋₆alkyl,        -   (c) —OH,        -   (d) —CN, or        -   (e) —O—C₁₋₆alkyl; and    -   R⁷ is selected from the group consisting of        -   (a) hydrogen, and        -   (b) —C₁₋₆alkyl,        -   wherein said alkyl is unsubstituted or substituted with one            or more            -   (i) halo,            -   (ii) —C₁₋₆alkyl,            -   (iii) —OH,            -   (iv) —CN, or            -   (v) —O—C₁₋₆alkyl;

-   R³ is selected from the group consisting of:

wherein R⁵ is C₁₋₆alkyl, C₂₋₆ alkenyl or C₂₋₆ alkynyl;

-   R^(6a), R^(6b), and R^(6c) are independently selected from the group    consisting of:    -   (1) hydrogen,    -   (2) halo,    -   (3) —C₁₋₆alkyl,    -   (4) —C₂₋₆ alkenyl,    -   (5) —C₂₋₆ alkynyl,    -   (6) —OH,    -   (7) —CN, and    -   (8) —O—C₁₋₆alkyl;-   R⁹ and R¹⁰ are independently selected from the group consisting of:    -   (1) hydrogen,    -   (2) —C₁₋₆alkyl,    -   (3) —C₂₋₆alkenyl, and    -   (4) —C₂₋₆ alkynyl,        or R⁹ and R¹⁰ are joined together with the nitrogen atom to        which they are attached to form a pyrrolidine ring, which is        optionally substituted with    -   (a) —C₁₋₆alkyl,    -   (b) —C₂₋₆alkenyl,    -   (c) —C₂₋₆ alkynyl,    -   (d) (CH₂)_(n)-phenyl, or    -   (e) (CH₂)_(n)-furanyl;    -   wherein said alkyl, phenyl and furanyl are unsubstituted or        substituted with one or more        -   i) halo,        -   ii) —C₁₋₆alkyl,        -   iii) —OH,        -   iv) —CN, or        -   v) —O—C₁₋₆alkyl; and-   R¹¹ is selected from the group consisting of    -   (1) —CH—,    -   (2) —O—, and    -   (3) —NH—,        provided that when R¹¹ is —CH— the dotted line forms a bond and        when R¹¹ is —O— or —NH— the dotted line is absent;-   R¹² is hydrogen, —C₁₋₆alkyl, C₂₋₆ alkenyl or C₂₋₆ alkynyl;-   m is 1 or 2;-   n is 0, 1, 2, 3 or 4;-   p is 1, 2, 3 or 4;    and pharmaceutically acceptable salts thereof.

In one embodiment of the compounds of formula (I), m is 1 and R¹ isphenyl, unsubstituted or substituted in one or two positions with halo,preferably with fluoro or chloro. In another embodiment, m is 2 and R¹is phenyl, unsubstituted or substituted in one or two positions withhalo, preferably with fluoro or chloro. In another embodiment, m is 1and R¹ is thiophenyl.

In another embodiment of the compounds of formula (I), R² isR⁴—S(O)₂N(R⁷)—, wherein R⁴ and R⁷ are each C₁₋₆alkyl, for example areeach methyl.

In another embodiment of the compounds of formula (I), R² is

In another embodiment of the compounds of formula (I), X is O.

In another embodiment of the compounds of formula (I), Y is CH.

An embodiment of the present invention is directed to compounds of theformula II:

wherein X, Y, R¹, R², R⁵, R^(6a), R^(6b), R^(6c) and m are as definedherein, and pharmaceutically acceptable salts thereof.

In one embodiment of the compounds of formula (II), R⁵ is methyl. Inanother embodiment of the compounds of formula (II), R^(6a) and R^(6b)are hydrogen and R^(6c) is fluoro.

In another embodiment of the compounds of formula (II), X is O.

In another embodiment of the compounds of formula (U), Y is CH.

Another embodiment of the present invention is directed to compounds ofthe formula (III):

wherein X, Y, R¹, R², R⁹, R¹⁰ and m are as defined herein.

In another embodiment of the compounds of formula (III), R⁹ and R¹⁰ arejoined together to form a pyrrolidine ring which is unsubstituted orsubstituted with —(CH₂)_(n)-furanyl wherein n is 0.

In another embodiment of the compounds of formula (III), X is O.

In another embodiment of the compounds of formula (III), Y is CH.

Another embodiment of the present invention is directed to compounds ofthe formula (IV):

wherein X, R¹, R², R¹¹, R¹² and m are as defined herein.

In an embodiment of the compounds of formula (IV), X is O.

In another embodiment of the compounds of formula (IV), Y is CH.

Another embodiment of the present invention includes a compound which isselected from the title compounds of the following Examples andpharmaceutically acceptable salts thereof.

As used herein, the term “alkyl,” by itself or as part of anothersubstituent, means a straight or branched chain hydrocarbon radicalhaving the number of carbon atoms designated (e.g., C₁₋₆ alkyl means analkyl group having from 1 to 6 carbon atoms). Exemplary alkyl groupsinclude methyl, ethyl, n-propyl, isopropyl, n-butyl, isobutyl,tert-butyl, pentyl, hexyl, and the like.

As used herein, the term “alkenyl,” by itself or as part of anothersubstituent, means a straight or branched chain hydrocarbon radicalhaving a single-carbon-carbon double bond and the number of carbon atomsdesignated (e.g., C₂₋₁₀ alkenyl means an alkenyl group having from twoto ten carbon atoms). Preferred alkenyl groups for use in the inventionare C₂₋₆ alkenyl groups, having from two to six carbon atoms. Exemplaryalkenyl groups include ethenyl and propenyl.

As used herein, the term “alkynyl” by itself or as part of anothersubstituent, means a straight or branched chain hydrocarbon radicalhaving a single carbon-carbon triple bond and the number of carbon atomsdesignated (e.g., C₂₋₁₀ alkynyl means an alkynyl group having from twoto ten carbon atoms). Preferred alkynyl groups for use in the inventionare C₂₋₆ alkynyl groups, having from two to six carbon atoms. Exemplaryalkynyl groups include ethynyl and propynyl.

The term “halo” or “halogen” includes fluoro, chloro, bromo and iodo.

The compounds of the instant invention have at least one asymmetriccenter. Additional asymmetric centers may be present depending upon thenature of the various substituents on the molecule. Compounds withasymmetric centers give rise to enantiomers (optical isomers),diastereomers (configurational isomers) or both, and it is intended thatall of the possible enantiomers and diastereomers in mixtures and aspure or partially purified compounds are included within the scope ofthis invention. The present invention is meant to encompass all suchisomeric forms of these compounds.

The independent syntheses of the enantiomerically or diastereomericallyenriched compounds, or their chromatographic separations, may beachieved as known in the art by appropriate modification of themethodology disclosed herein. Their absolute stereochemistry may bedetermined by the x-ray crystallography of crystalline products orcrystalline intermediates that are derivatized, if necessary, with areagent containing an asymmetric center of known absolute configuration.

If desired, racemic mixtures of the compounds may be separated so thatthe individual enantiomers are isolated. The separation can be carriedout by methods well known in the art, such as the coupling of a racemicmixture of compounds to an enantiomerically pure compound to form adiastereomeric mixture, followed by separation of the individualdiastereomers by standard methods, such as fractional crystallization orchromatography. The coupling reaction is often the formation of saltsusing an enantiomerically pure acid or base. The diastereomericderivatives may then be converted to the pure enantiomers by cleavage ofthe added chiral residue. The racemic mixture of the compounds can alsobe separated directly by chromatographic methods utilizing chiralstationary phases, which methods are well known in the art.

Alternatively, any enantiomer of a compound may be obtained bystereoselective synthesis using optically pure starting materials orreagents of known configuration by methods well known in the art.

The compounds of the present invention are prepared by the methodsoutlined in Scheme 1, below.

In Scheme 1, an alkyl serine derivative of type 1 is converted to thecorresponding alcohol 2 which in turn is protected as the Boc derivative3. Deprotonation and alkylation of diethyl2-[N-(tert-butoxycarbonyl)-amino]malonate 4 with various alkyl or benzylbromides, followed by reduction provides an alternative to thepreparation of diols of type 3. Boc removal then provides an alternativeroute to 2. The alkyl serine derivative of type 1 is esterified withMeOH, and protected with Boc, and the hydroxyl group is converted to thecorresponding azide which is reduced to afford an amine of type 5.

In Scheme 2, below, dimethyl 5-aminoisophthalate 6 is converted tobromide 7 via a 7-step sequence involving sulfonylation, alkylation,hydrolysis, amine coupling, hydrolysis, reduction, and bromination.Deprotonation of diol 2 followed by alkylation with bromide 7 givesaccess to derivative 8. Alternatively, diol 3 can be alkylated withbromide 7 in the presence of silver triflate to provide 8 after Bocremoval. Alkylation of amine 5 with bromide 7, followed by esterreduction and Boc removal, affords amine 9.

In Schemes 2-4, NR′R″ represent the R³ groups (a) or (b), describedabove.

Scheme 3 illustrates the preparation of bromide of type 11 from dimethyl5-iodoisophthalate 10 via Pd coupling, hydrolysis, reduction andbromination. Alkylation of diol 3 with bromide 11, followed byhydrolysis, amine coupling and Boc removal gives access to ether 12.

Scheme 4 illustrates the preparation of bromide of type 15 that is thencoupled to diol 2 or 3, as described in scheme 2. Installation of bothside chains using PdO coupling methodology followed by reduction of theester moiety and subsequent bromination provides 15.

Scheme 5 illustrates the preparation of bromides of type 19 and 22 thatare then coupled to diol 2 or 3, as described in scheme 2. Phenol 16 isalkylated and the methyl ester is converted to a bromomethylfunctionality giving access to intermediate 17. The cyano-cycloalkylgroup is introduced via TMS-CN and the necessary dibromoalkane.Subsequent cyclopropanation provides 18 that is converted to bromide 19.The preparation of bromide 22 relies on similar methodology regardingthe R¹²-bearing side chain and a Curtius rearrangement for theintroduction of R⁷NSO₂R⁴.

Scheme 6 illustrates two alternative preparations of bromide of type 23that is then coupled to diol 2 or 3, as described in scheme 2. The firstpreparation relies on conversion of the methyl ester to an aldehyde anda Wittig coupling to install the R⁵-bearing alkene. The secondpreparation is based on an indenium/palladium coupling strategy.

The term “substantially pure” means that the isolated material is atleast 90% pure, and preferably 95% pure, and even more preferably 99%pure as assayed by analytical techniques known in the art.

The term “pharmaceutically acceptable salts” refers to salts preparedfrom pharmaceutically acceptable non-toxic bases or acids includinginorganic or organic bases and inorganic or organic acids. Salts derivedfrom inorganic bases include aluminum, ammonium, calcium, copper,ferric, ferrous, lithium, magnesium, manganic salts, manganous,potassium, sodium, zinc, and the like. Particularly preferred are theammonium, calcium, magnesium, potassium, and sodium salts. Salts in thesolid form may exist in more than one crystal structure, and may also bein the form of hydrates. Salts derived from pharmaceutically acceptableorganic non-toxic bases include salts of primary, secondary, andtertiary amines, substituted amines including naturally occurringsubstituted amines, cyclic amines, and basic ion exchange resins, suchas arginine, betaine, caffeine, choline, N,N-dibenzylethylene-diamine,diethylamine, 2-diethylaminoethanol, 2-dimethylaminoethanol,ethanolamine, ethylenediamine, N-ethyl-morpholine, N-ethylpiperidine,glucamine, glucosamine, histidine, hydrabamine, isopropylamine, lysine,methylglucamine, morpholine, piperazine, piperidine, polyamine resins,procaine, purines, theobromine, triethylamine, trimethylamine,tripropylamine, tromethamine, and the like. When the compound of thepresent invention is basic, salts may be prepared from pharmaceuticallyacceptable non-toxic acids, including inorganic and organic acids. Suchacids include acetic, benzenesulfonic, benzoic, camphorsulfonic, citric,ethanesulfonic, fumaric, gluconic, glutamic, hydrobromic, hydrochloric,isethionic, lactic, maleic, malic, mandelic, methanesulfonic, mucic,nitric, pamoic, pantothenic, phosphoric, succinic, sulfuric, tartaric,p-toluenesulfonic, trifluoracetic acid, and the like. Particularlypreferred are citric, hydrobromic, hydrochloric, maleic, phosphoric,sulfuric, fumaric, tartaric and trifluoracetic acids.

The present invention is directed to the use of the compounds disclosedherein as inhibitors of β-secretase enzyme activity or β-site amyloidprecursor protein-cleaving enzyme (“BACE”) activity, in a patient orsubject such as a mammal in need of such inhibition, comprising theadministration of an effective amount of the compound. The terms“β-secretase enzyme,” “β-site amyloid precursor protein-cleavingenzyme,” and “BACE” are used interchangably in this specification. Inaddition to humans, a variety of other mammals can be treated accordingto the method of the present invention.

The present invention is further directed to a method for themanufacture of a medicament or a composition for inhibiting β-secretaseenzyme activity in humans and animals comprising combining a compound ofthe present invention with a pharmaceutical carrier or diluent.

The compounds of the present invention have utility in treatingAlzheimer's disease. For example, the compounds may be useful intreating dementia of the Alzheimer's type, including early stage,intermediate stage or late stage dementia of the Alzheimer's type. Thecompounds may also be useful in treating diseases mediated by abnormalcleavage of amyloid precursor protein (also referred to as APP), andother conditions that may be treated or prevented by inhibition ofβ-secretase. Such conditions include mild cognitive impairment, Trisomy21 (Down Syndrome), cerebral amyloid angiopathy, degenerative dementia,Hereditary Cerebral Hemorrhage with Amyloidosis of the Dutch-Type(HCHWA-D), Creutzfeldt-Jakob disease, prion disorders, amyotrophiclateral sclerosis, progressive supranuclear palsy, head trauma, stroke,Down syndrome, pancreatitis, inclusion body myositis, other peripheralamyloidoses, diabetes and atherosclerosis.

The subject or patient to whom the compounds of the present invention isadministered is generally a human being, male or female, in whominhibition of β-secretase enzyme activity is desired, but may alsoencompass other mammals, such as dogs, cats, mice, rats, cattle, horses,sheep, rabbits, monkeys, chimpanzees or other apes or primates, forwhich inhibition of β-secretase enzyme activity or treatment of theabove noted disorders is desired.

The compounds of the present invention may be used in combination withone or more other drugs in the treatment of diseases or conditions forwhich the compounds of the present invention have utility, where thecombination of the drugs together are safer or more effective thaneither drug alone. Additionally, the compounds of the present inventionmay be used in combination with one or more other drugs that treat,prevent, control, ameliorate, or reduce the risk of side effects ortoxicity of the compounds of the present invention. Such other drugs maybe administered, by a route and in an amount commonly used therefor,contemporaneously or sequentially with the compounds of the presentinvention. Accordingly, the pharmaceutical compositions of the presentinvention include those that contain one or more other activeingredients, in addition to the compounds of the present invention. Thecombinations may be administered as part of a unit dosage formcombination product, or as a kit or treatment protocol wherein one ormore additional drugs are administered in separate dosage forms as partof a treatment regimen.

Examples of combinations of the compounds of the present invention withother drugs in either unit dose or kit form include combinations withanti-Alzheimer's agents, for example other beta-secretase inhibitors;gamma-secretase inhibitors; HMG-CoA reductase inhibitors; NSAID'sincluding ibuprofen; vitamin E; anti-amyloid antibodies, includinghumanized monoclonal antibodies; CB-1 receptor antagonists or CB-1receptor inverse agonists; antibiotics such as doxycycline and rifampin;N-methyl-D-aspartate (NMDA) receptor antagonists, such as mematine;cholinesterase inhibitors such as galantamine, rivastigmnine, donepeziland tacrine; growth hormone secretagogues such as ibutamoren, ibutamorenmesylate and capromorelin; histamine H₃ antagonists; AMPA agonists,PDEIV inhibitors; GABA_(A) inverse agonists; neuronal nicotinicagonists; or other drugs that affect receptors or enzymes that eitherincrease the efficacy, safety, convenience, or reduce unwanted sideeffects or toxicity of the compounds of the present invention. Theforegoing list of combinations is illustrative only and not intended tobe limiting in any way.

The term “composition” as used herein is intended to encompass a productcomprising specified ingredients in predetermined amounts orproportions, as well as any product which results, directly orindirectly, from combination of the specified ingredients in thespecified amounts. This term in relation to pharmaceutical compositionsis intended to encompass a product comprising one or more activeingredients, and an optional carrier comprising inert ingredients, aswell as any product which results, directly or indirectly, fromcombination, complexation or aggregation of any two or more of theingredients, or from dissociation of one or more of the ingredients, orfrom other types of reactions or interactions of one or more of theingredients. In general, pharmaceutical compositions are prepared byuniformly and intimately bringing the active ingredient into associationwith a liquid carrier or a finely divided solid carrier or both, andthen, if necessary, shaping the product into the desired formulation. Inthe pharmaceutical composition the active object compound is included inan amount sufficient to produce the desired effect upon the process orcondition of diseases. Accordingly, the pharmaceutical compositions ofthe present invention encompass any composition made by admixing acompound of the present invention and a pharmaceutically acceptablecarrier.

Pharmaceutical compositions intended for oral use may be preparedaccording to any method known to the art for the manufacture ofpharmaceutical compositions and such compositions may contain one ormore agents selected from the group consisting of sweetening agents,flavoring agents, coloring agents and preserving agents in order toprovide pharmaceutically elegant and palatable preparations. Tabletscontain the active ingredient in admixture with non-toxicpharmaceutically acceptable excipients which are suitable for themanufacture of tablets. These excipients may be for example, inertdiluents, such as calcium carbonate, sodium carbonate, lactose, calciumphosphate or sodium phosphate; granulating and disintegrating agents,for example, corn starch, or alginic acid; binding agents, for examplestarch, gelatin or acacia, and lubricating agents, for example magnesiumstearate, stearic acid or talc. The tablets may be uncoated or they maybe coated by known techniques to delay disintegration and absorption inthe gastrointestinal tract and thereby provide a sustained action over alonger period.

Compositions for oral use may also be presented as hard gelatin capsuleswherein the active ingredient is mixed with an inert solid diluent, forexample, calcium carbonate, calcium phosphate or kaolin, or as softgelatin capsules wherein the active ingredient is mixed with water or anoil medium, for example peanut oil, liquid paraffin, or olive oil.

Other pharmaceutical compositions include aqueous suspensions, whichcontain the active materials in admixture with excipients suitable forthe manufacture of aqueous suspensions. In addition, oily suspensionsmay be formulated by suspending the active ingredient in a vegetableoil, for example arachis oil, olive oil, sesame oil or coconut oil, orin a mineral oil such as liquid paraffin. Oily suspensions may alsocontain various excipients. The pharmaceutical compositions of theinvention may also be in the form of oil-in-water emulsions, which mayalso contain excipients such as sweetening and flavoring agents.

The pharmaceutical compositions may be in the form of a sterileinjectable aqueous or oleagenous suspension, which may be formulatedaccording to the known art, or may be administered in the form ofsuppositories for rectal administration of the drug.

The compounds of the present invention may also be administered byinhalation, by way of inhalation devices known to those skilled in theart, or by a transdermal patch.

By “pharmaceutically acceptable” it is meant the carrier, diluent orexcipient must be compatible with the other ingredients of theformulation and not deleterious to the recipient thereof.

The terms “administration of” or “administering a” compound should beunderstood to mean providing a compound of the invention to theindividual in need of treatment in a form that can be introduced intothat individual's body in a therapeutically useful form andtherapeutically useful amount, including, but not limited to: oraldosage forms, such as tablets, capsules, syrups, suspensions, and thelike; injectable dosage forms, such as IV, IM, or IP, and the like;transdermal dosage forms, including creams, jellies, powders, orpatches; buccal dosage forms; inhalation powders, sprays, suspensions,and the like; and rectal suppositories.

The terms “effective amount” or “therapeutically effective amount” meansthe amount of the subject compound that will elicit the biological ormedical response of a tissue, system, animal or human that is beingsought by the researcher, veterinarian, medical doctor or otherclinician. As used herein, the term “treatment” refers to the treatmentof the mentioned conditions, particularly in a patient who demonstratessymptoms of the disease or disorder.

As used herein, the term “treating” means any administration of acompound of the present invention and includes (1) inhibiting thedisease in an animal that is experiencing or displaying the pathology orsymptomatology of the disease (i.e., arresting further development ofthe pathology and/or symptomatology), or (2) ameliorating the disease inan animal that is experiencing or displaying the pathology ofsymptomatology of the disease (i.e., reversing theathology and/orsymptomatology). The term “controlling” includes preventing treating,eradicating, ameliorating or otherwise reducing the severity of thecondition being controlled.

The compositions containing compounds of the present invention mayconveniently be presented in unit dosage form and may be prepared by anyof the methods well known in the art of pharmacy. The term “unit dosageform” is taken to mean a single dose wherein all active and inactiveingredients are combined in a suitable system, such that the patient orperson administering the drug to the patient can open a single containeror package with the entire dose contained therein, and does not have tomix any components together from two or more containers or packages.Typical examples of unit dosage forms are tablets or capsules for oraladministration, single dose vials for injection, or suppositories forrectal administration. This list of unit dosage forms is not intended tobe limiting in any way, but merely to represent typical examples in thepharmacy arts of unit dosage forms.

The compositions containing compounds of the present invention mayconveniently be presented as a kit, whereby two or more components,which may be active or inactive ingredients, carriers, diluents, and thelike, are provided with instructions for preparation of the actualdosage form by the patient or person administering the drug to thepatient. Such kits may be provided with all necessary materials andingredients contained therein, or they may contain instructions forusing or making materials or components that must be obtainedindependently by the patient or person administering the drug to thepatient.

When treating Alzheimer's disease or other diseases for which compoundsof the present invention are indicated, generally satisfactory resultsare obtained when the compounds of the present invention areadministered at a daily dosage of from about 0.1 mg to about 100 mg perkg of animal body weight, preferably given as a single daily dose or individed doses two to six times a day, or in sustained release form. Thetotal daily dosage is from about 1.0 mg to about 2000 mg, preferablyfrom about 0.1 mg to about 20 mg per kg of body weight. In the case of a70 kg adult human, the total daily dose will generally be from about 7mg to about 1,400 mg. This dosage regimen may be adjusted to provide theoptimal therapeutic response. The compounds may be administered on aregimen of 1 to 4 times per day, preferably once or twice per day.

Specific dosages of the compounds of the present invention, orpharmaceutically acceptable salts thereof, for administration include 1mg, 5 mg, 10 mg, 30 mg, 100 mg, 300 mg and 500 mg. Pharmaceuticalcompositions of the present invention may be provided in a formulationcomprising about 0.5 mg to 1000 mg active ingredient; more preferablycomprising about 0.5 mg to 500 mg active ingredient; or 0.5 mg to 250 mgactive ingredient; or 1 mg to 100 mg active ingredient. Specificpharmaceutical compositions may comprise about 1 mg, 5 mg, 10 mg, 30 mg,100 mg, 300 mg and 500 mg of active ingredient.

It will be understood, however, that the specific dose level andfrequency of dosage for any particular patient may be varied and willdepend upon a variety of factors including the activity of the specificcompound employed, the metabolic stability and length of action of thatcompound, the age, body weight, general health, sex, diet, mode and timeof administration, rate of excretion, drug combination, the severity ofthe particular condition, and the host undergoing therapy.

The utility of the compounds in accordance with the present invention asinhibitors of β-secretase enzyme activity may be demonstrated bymethodology known in the art. Enzyme inhibition is determined asfollows.

FRET Assay: A homogeneous end point fluorescence resonance energytransfer (FRET) assay is employed with the substrate([TAMRA-5-CO-EEISEVNLDAEF-NHQSY] QFRET), which is cleaved by BACE 1 torelease the fluorescence from TAMRA. The Km of the substrate is notdetermined due to the limit of solubility of the substrate. A typicalreaction contains approximately 30 nM enzyme, 1.25 μM of the substrate,and buffer (50 mM NaOAc, pH 4.5, 0.1 mg/ml BSA, 0.2% CHAPS, 15 mM EDTAand 1 mM deferoxamine) in a total reaction volume of 100 μl. Thereaction is proceeded for 30 min and the liberation of TAMRA fragment ismeasured in a 96-well plate LJL Analyst AD using an excitationwavelength of 530 nm and an emission wavelength of 580 nm. Under theseconditions, less than 10% of substrate is processed by BACE 1. Theenzyme used in these studies was soluble (transmembrane domain andcytoplasmic extension excluded) human protein produced in a baculovirusexpression system. To measure the inhibitory potency of compounds,solutions of inhibitor in DMSO (four concentrations of the inhibitorswere prepared: 1 mM, 100 μM, 10 μM, 1 μM) were included in the reactionsmixture (final DMSO concentration is 0.8%). All experiments wereconducted at room temperature using the standard reaction conditionsdescribed above. To determine the IC₅₀ of the compound, competitiveequation V0/Vi=1+[I]/[IC50] was used to predict the inhibitory potencyof the compounds. The errors in reproducing the dissociation constantsare typically less than two-fold.

HPLC assay: A homogeneous end point HPLC assay is employed with thesubstrate (coumarin-CO-REVNFEVEFR), which is cleaved by BACE 1 torelease the N-terminal fragment attached with coumarin. The Km of thesubstrate is greater than 100 μM and can not be determined due to thelimit of solubility of the substrate. A typical reaction containsapproximately 2 nM enzyme, 1.0 μM of the substrate, and buffer (50 mMNaOAc, pH 4.5, 0.1 mg/ml BSA, 0.2% CHAPS, 15 mM EDTA and 1 mMdeferoxamine) in a total reaction volume of 100 μl. The reaction isproceeded for 30 min and is stopped by the addition of 25 μL of 1 MTris-HCl, pH 8.0. The resulting reaction mixture was loaded on the HPLCand the product was separated from substrate with 5 min linear gradient.Under these conditions, less than 10% of substrate is processed byBACE 1. The enzyme used in these studies was soluble (transmembranedomain and cytoplasmic extension excluded) human protein produced in abaculovirus expression system. To measure the inhibitory potency forcompounds, solutions of inhibitor in DMSO (12 concentrations of theinhibitors were prepared and the concentration rage was dependent on thepotency predicted by FRET) were included in the reaction mixture (finalDMSO concentration is 10%). All experiments were conducted at roomtemperature using the standard reaction conditions described above. Todetermine the IC₅₀ of the compound, four parameters equation is employedfor curve fitting. The errors in reproducing the dissociation constantsare typically less than two-fold.

In particular, the compounds of the following examples had activity ininhibiting the beta-secretase enzyme in the aforementioned assay,generally with an IC50 from about 1 nM to 100 μM. Such a result isindicative of the intrinsic activity of the compounds in use asinhibitors of beta-secretase enzyme activity.

Several methods for preparing the compounds of this invention areillustrated in the following Schemes and Examples. Starting materialsare made according to procedures known in the art or as illustratedherein. The following examples are provided so that the invention mightbe more fully understood. These examples are illustrative only andshould not be construed as limiting the invention in any way.

The following abbreviations are used throughout the text:

Ar: aryl

Ph: phenyl

Me: methyl

Et: ethyl

Ac: acetyl

t-bu: tert-butyl

Pyr: pyridine

TFA: trifluoroacetic acid

DMF: N,N′-dimethyl formamide

THF: tetrahydrofuran

LAH: lithium aluminum hydride

TEA: trifluoroacetic acid

DMSO: dimethylsulfoxide

HPLC: high performance liquid chromatography

EDTA: ethylene diamine tetraacetic acid

Boc: tert-butyloxy carbonyl

rac: racemic

DIBAL: diisobutylaluminium hydride

BOP: Benzotriazol-1-yloxy-tris(dimethylamino)phosphoniumhexafluorophosphate

DEAD: diethylazole dicarboxylate

Intermediate I: 2-amino-2-benzylpropane-1,3-diol

To a solution of rac-benzylserine (4.00 g, 20.49 mmol) in 60 mL THF atroom temperature was added NaBH₄ (2.71 g 71.71 mmol) in one portion. Thesolution was fitted with a reflux condenser and cooled to 0° C. Iodine(7.80 g, 30.73 mmol) in 20 mL THF was added dropwise via cannula. Afterthe addition was complete, the reaction was heated to reflux for 15 h.The reaction was then cooled to 0° C. and quenched by the addition ofmethanol until the bubbling subsided. The residue was acidified by theaddition of 6N HCl until the pH was below one, then concentrated toafford 2-amino2-benzylpropane-1,3-diol contaminated with inorganicresidue. The unpurified reaction mixture was redissolved in methanol andfiltered through a pad of celite (rinsing copiously with fresh methanol)to remove some of the insoluble inorganic residue. After concentration,further purification was accomplished using ion exchange chromatography(SCX cartridge) to afford 2-amino2-benzylpropane-1,3-diol I as a whitesolid. ¹H NMR (400 M, d₄-MeOH) δ 7.34-7.25 (m, 5H), 3.52 (s, 4H), 3.00(s, 2H).

Intermediate II:tert-butyl[1-benzyl-2-hydroxy-1-(hydroxymethyl)ethyl]carbamate

To a solution of 2-amino-2-benzylpropane-1,3-diol I (0.901 g, 4.971mmol) in 50 mL THF was added ditert-butyl dicarbonate (1.139 g, 5.22nmol). After 14 h at room temperature, the reaction was concentrated,and purified by flash chromatography (40 g silica, 0→10% MeOH/CH₂Cl₂) toafford tert-butyl[1-benzyl-2-hydroxy-1-(hydroxymethyl)ethyl]carbamate IIas a white solid. ¹H NMR (400 MHz, d₄-MeOH) δ 7.25-7.15 (m, 5H), 3.59(d, J=11.0 Hz, 2H), 3.54 (d, J=11.0 Hz, 2H), 2.98 (s, 2H), 1.449 (s,911).

Intermediate III: tert-butyl2-hydroxy-1-(hydroxymethyl)-1-(thien-3-ylmethyl)ethylcarbamate

Step A: Alkylation

To a solution of diethyl 2-[N-(tert-butoxycarbonyl)-amino]malonate (1.44mL, 5.65 mmol) in 30 mL EtOH cooled to 0° C. was added sodium ethoxide(2.22 mL, 5.93 mmol, 21% in EtOH). The reaction mixture was stirred at0° C. for 5 min and 3-bromomethyl-thiophene (1 g, 5.65 mmol) in 10 mLEtOH was added dropwise. The reaction mixture was allowed to warm toroom temperature over 14 h. The reaction mixture was quenched withwater, and extracted with EtOAc. The organic layer was washed withbrine, dried over sodium sulfate, concentrated in vacuo, and purified byflash chromatography (120 g silica, 0→25% EtOAc/hexanes) to afforddiethyl 2,2-[N-(tert-butoxycarbonyl)-amino]-(thien-3-ylmethyl)malonateas a colorless oil. ¹H NMR (400 MHz, CDCl₃) δ 7.22 (dd, J=4.8 Hz, 2.5Hz, 1H), 6.94 (d, J=2.5 Hz, 1H), 6.81 (d, J=4.8 Hz, 1H), 5.82 (br s,1H), 4.34-4.14 (m, 4H), 3.66 (s, 2H), 1.47 (s, 9H), 1.27 (t, J=7.2 Hz,6H).

Step B: Reduction

A solution of diethyl2,2-[N-(tert-butoxycarbonyl)-amino]-(thien-3-ylmethyl)malonate (1.52 g,12.69 mmol) in 10 mL diethyl ether was added dropwise to LAH (12.7 mL,12.7 mmol, 1M in diethyl ether) in 20 mL diethyl ether cooled to 0° C.The reaction mixture was stirred at 0° C. for 1.25 h. The reactionmixture was carefully quenched with water (0.48 mL), 15% NaOH (0.48 mL),water (1.45 mL), stirred vigorously at room temperature for 5 min,diluted with THF, filtered on cellite, rinsed with THF, dried oversodium sulfate, concentrated in vacuo, and purified by flashchromatography (120 g silica, 25→50% EtOAc/hexanes) to afford tert-butyl2-hydroxy-1-(hydroxymethyl)-1-(thien-3-ylmethyl)ethylcarbamate III as athick colorless oil. ¹H NMR (400 MHz, CD₃OD) δ 7.29 (dd, J=4.8 Hz, 3.2Hz, 1H), 7.08 (dd, J=3.2 Hz, 0.8 Hz, 1), 6.98 (dd, J=4.8, 0.8 Hz Hz,1H), 3.61 (A of AB, d, J=20.0 Hz, 2H), 3.55 (B of AB, d, J=20.0 Hz, 2H),3.04 (s, 2H), 1.47 (s, 9H).

Intermediate IV: methylalpha-(aminomethyl)-N-(tert-butoxycarbonyl)phenylalaninate

Step A: Hydroxy to Azide Conversion

To a solution of methylN-(tert-butoxycarbonyl)-alpha-(hydroxymethyl)phenylalaninate (0.5 g,1.62 mmol, prepared from benzylserine via esterification in MeOH and Bocinstallation (or according to A. Kozikowski et al, Bioorg. Med. Chem.Lett. 1998, 8, 447-452) in 10 mL THF was added hydrazoic acid (1.2 mL,2.42 mmol, 2M in benzene), triphenylphosphine (0.42 g, 1.62 mmol), anddiethylazodicarboxylate (0.28 mL, 1.78 mmol) in 5 mL THF dropwise. Thereaction mixture was stirred at room temperature for 16 h, concentratedin vacuo and purified by flash chromatography (90 g silica, 0→20%EtOAc/hexanes) to affordN-(tert-butoxycarbonyl)-alpha-azidomethyl)phenylalaninate. ¹H NMR (400MHz, CDCl₃) δ 7.30-7.20 (m, 3H), 7.05-6.98 (m, 2H), 5.47 (br s, 1H),4.33 (d, J=12.4 Hz, 1H), 3.78 (s, 3H), 3.61 (d, J=12.4 Hz, 1H), 3.51 (d,J=13.2 Hz, 1H), 2.96 (d, J=13.2 Hz, 1H), 1.48 (s, 9H).

Step B: Hydrogenation

To a solution ofN-(tert-butoxycarbonyl)-alpha-azidomethyl)phenylalaninate (144 mg, 0.43mmol) in 20 mL MeOH, purged with argon was added 10% Pd/C (15 mg) andthe system was flushed with hydrogen. The reaction mixture was stirredat room temperature under 1 atm of hydrogen for 1 h. Filtration oncellite, rinsing with MeOH and concentration in vacuo provided methylalpha-(aminomethyl)-N-(tert-butoxycarbonyl)phenylalaninate IV. ¹H NMR(400 MHz, CDCl₃) δ 7.30-7.20 (m, 3H), 7.08-7.02 (m, 2H), 5.45 (br s,1H), 3.78 (s, 3H), 3.56 (br d, J=13.6 Hz, 1H), 3.50 (br d, J=13.2 Hz,1H), 3.10 (d, J=13.2 Hz, 1H), 3.06 (d, J=13.6 Hz, 1H), 1.48 (s, 9H),1.07 (br s, 1H).

Intermediate A:3-(bromomethyl)-N-[(1R)-1-(4-fluorophenyl)ethyl]-5-[methyl(methylsulfonyl)amino]benzamide

Step A: Sulfonylation

To a stirred slurry of dimethyl 5-aminoisophthalate (5.0 g, 23.90 mmol)in 100 mL CH₂Cl₂/pyridine (3:1) at 0° C. was added methanesulfonylchloride (1.85 mL, 23.90 mmol). The resulting mixture was stirred for 4h at room temperature. The solvent was removed in vacuo and ethylacetate(100 mL) was added resulting in precipitate formation. The product wascollected by filtration to give the sulfonamide as a white solid. 1H NMR(DMSO_(d6)) δ 8.15 (s, 1H), 8.02 (s, 2H), 3.89 (s, 6H), 3.02 (s, 3H)LCMS [M-OCH₃]+=256.16.

Step B: Methylation

To a solution of sodium hydride (0153 g, 3.83 mmol, 60% oil dispersion)in 10 mL DMF was added sulfonamide (1.0 g, 3.48 mmol) from step Afollowed by methyl iodide (0.43 mL, 6.97 mmol). After 1 hr the reactionwas quenched with H₂O (100 mL) and extracted with EtOAc (3×50 mL). Theorganic extracts were dried over MgSO₄ and evaporated to give theproduct. ¹H NMR (DMSO_(d6)) δ 8.40 (s, 1H), 8.19 (s, 2H), 3.91 (s, 6H),3.34 (s, 3H), 3.01 (s, 3H). LCMS [M+H]=302.15.

Step C: Hydrolysis

Diester (1.03 g, 3.38 mmol) from step B was dissolved in 50 mL THF:MeOH(1:1) and cooled to 0° C. 1N NaOH (3.38 mL, 3.38 mmol) was added and thereaction was allowed to warm to RT over 8 hours. The solution wasacidified with 1N HCl (30 mL) and extracted with EtOAc (3×50 mL). Thecombined organic extracts were washed with brine and dried over MgSO₄,filtered and concentrated in vacuo. Purification on silica gel (5%MeOH/CHCl₃ containing 1% HOAc) gave the mono acid. ¹H NMR (DMSO_(d6)) δ8.30 (s, 1H), 8.10 (s, 2H), 3.84 (s, 3H), 3.27 (s, 3H), 2.94 (s, 3H).LCMS (M+H)=288.16.

Step D: Amine Coupling

A solution containing 0.133 g (0.46 mmol) of the monoacid from step C in5 mL CH₂Cl₂, BOP reagent (0.235 g, 0.55 mmol),(R)-(+)-α-methylbenzylamine (0.071 mL, 0.55 mmol), and diisopropylamine(0.24 mL, 1.39 mmol) was stirred at ambient temperature for 1 h.Evaporation of the solvent and column chromatography on silica gel (90%EtOAc/Hexanes) afforded the benzyl amide. ¹H NMR (CDCl₃) δ 8.26 (s, 1H),8.17 (s, 1H), 8.06 (s, 1H), 7.31 (m, 5H), 6.50 (d, J=7.1 Hz, 1H), 5.33(q, J=7.1 Hz, 1H), 3.96 (s, 3H), 3.37 (s, 3H), 2.88 (s, 3H), 1.64 (d,J=7.0 Hz, 3H). LCMS (M+H)=391.20.

Step E: Hydrolysis

To 0.171 g (0.438 mmol) of the benzyl amide from step D in 10 mLTHF:MeOH (1:1) was added 2 N NaOH (0.66 mL, 1.32 mmol). The solution washeated to 50° C. for 1 h. After cooling the solution was acidified bythe addition of 1 N HCl (20 mL) and extracted with EtOAc (3×30 mL). Thecombined organic extracts were dried over MgSO4, filtered, andconcentrated in vacuo to yield the desired carboxylic acid. 1H NMR(CDCl₃) δ 8.22 (t, 1H), 8.11 (m, 1H), 8.06 (m, 1H), 7.34 (m, 5H), 6.47(d, J=7.1 Hz, 1H), 5.33 (m, 1H), 3.37 (s, 3H), 2.87 (s, 3H), 1.64 (d,J=7.0 Hz, 3H). LCMS (M+H)=377.2.

Step E: Reduction

To a solution of acid from step D (800 mg, 2.0 mmol) in 20 mL THF cooledto 0° C. was added borane (6.0 ml, 6.0 mmol, 1 M in THF) dropwise. Thereaction mixture was stirred at 0° C. for 20 min and at room temperaturefor 1 h 45 min. The reaction mixture was quenched with MeOH and stirredat room temperature for 16 h, and concentrated in vacuo. The residue wastaken in EtOAc, washed with water, brine, dried over sodium sulfate andconcentrated in vacuo to provide crude alcohol which was brominated asis in step F.

Step F: Bromination

To a solution of crude alcohol from step E (355 mg, 0.93 mmol) andcarbon tetrabromide (0.4 g, 1.2 mmol) in 4.6 mL 1:1 CH₃CN:CH₂Cl₂ wasadded triphenylphosphine (0.29 g, 1.1 mmol) in 4.6 mL 1:1 CH₃CN:CH₂Cl₂dropwise. After stirring at room temperature for 45 min, two additionalbatches of carbon tetrabromide and triphenyl phosphine (200 mg/15 mg and20 mg/15 mg) were added at 30 min intervals, until the reaction appearedcomplete by LC/MS analysis. The reaction mixture was concentrated andpurified by flash chromatography (40 g silica, 25→60% EtOAc/hexanes) toafford 220 mg of3-(bromomethyl)-N-[(1R)-1-(4-fluorophenyl)ethyl]-5-[methyl(methylsulfonyl)amino]benzamideA. 1H NMR (400 MHz, CDCl₃) δ 7.73 (s, 1H), 7.65 (s, 1H), 7.57 (s, 1H),7.32-7.40 (m, 2H), 7.09-7.01 (m, 2H), 6.32 (d, J=7.6 Hz, 1H), 5.36-5.24(m, 1H), 4.50 (s, 2H), 3.36 (s, 3H), 2.78 (s, 3H), 1.62 (d, J=6.5 Hz,3H).

Intermediate B: methyl5-(bromomethyl)-2′-cyano-1,1′-biphenyl-3-carboxylate

To a solution of dimethyl 5-iodoisophthalate (13 g, 40.6 mmol) in 100 mLTHF was added 2-cyano-phenyl zinc bromide (97.5 mL, 48.7 mmol, 0.5 M THFand tetrakis(triphenylphosphine)palladium (214 mg, 0.2 mmol) and thereaction mixture was stirred at room temperature for 2 h. Theprecipitated solid was filtered, the filtrate was diluted with MeOH toprovide after filtration of a second crop dimethyl5-(2-cyanophenyl)isophthalate which was hydrolyzed to the correspondingmonoacid 2′-cyano-5-(methoxycarbonyl)-1,1′-biphenyl-3-carboxylic acidfollowing a similar procedure as described in intermediate Apreparation, step C. ¹H NMR (400 MHz, d₆-DMSO) δ 13.55 (br s, 1H),8.60-8.55 (m, 1H), 8.38-8.31 (m, 2H), 8.02 (d, J=8.3 Hz, 1H), 7.85 (td,J=8.3 Hz, 1.5 Hz 1H), 7.75 (d, J=8.3 Hz, 1H), 7.66 (td, J=8.3 Hz, 1.5 Hz1H), 3.93 (s, 3H).

Reduction with borane and bromination using a similar procedure asdescribed in intermediate A preparation, steps E and F affordedintermediate B: methyl5-(bromomethyl)-2′-cyano-1,1′-biphenyl-3-carboxylate. ¹H NMR (400 MHz,CDCl₃) δ 8.12-8.18 (m, 2H), 7.83-7.77 (m, 2H), 7.69 (td, J=8.5 Hz, 1.4Hz 1H), 7.55 (d, J=8.5 Hz, 1H), 7.51 (td, J=8.5 Hz, 1.4 Hz 1H), 4.59 (s,2H), 3.96 (s, 3H).

Example 13-[(2-amino-2-benzyl-3-hydroxypropoxy)methyl]-N-[(1R)-1-(4-fluorophenyl)ethyl]-5-[methyl(methylsulfonyl)amino]benzamide

To a solution of intermediate I 2-amino-2-benzylpropane-1,3-diol (0.09g, 0.50 mmol) in 2 mL DMF cooled to 0° C. was added sodiumhexamethyldisylazide (0.5 mL, 0.50 mmol, 1 M in THF). The reactionmixture was stirred at 0° C. for 5 min and intermediate A3-(bromomethyl)-N-[(1R)-1-(4-fluorophenyl)ethyl]-5-[methyl(methylsulfonyl)amino]benzamide(0.1 g, 0.23 mmol) in 1 mL DMF was added dropwise. The reaction mixturewas stirred at 0° C. for 0.5 h, quenched with water, extracted withEtOAc, washed with aq LiCl (3×), dried over sodium sulfate, concentratedin vacuo, and purified by flash chromatography (40 g silica, 0→8% (10%NH₄OH in MeOH)/CH₂Cl₂) to afford3-[(2-amino-2-benzyl-3-hydroxypropoxy)methyl]-N-[(1R)-1-(4-fluorophenyl)ethyl]-5-[methyl(methylsulfonyl)amino]benzamide.¹H NMR (400 MHz, d₄-MeOH) δ 7.80 (s, 2H), 7.64 (s, 1H), 7.44-7.40 (m,2H), 7.26-7.20 (m, 5H), 7.07-7.02 (m, 2H), 5.24 (q, J=7.0 Hz, 1H), 4.61(s, 2H), 3.48 (A of AB, d, J=10.4 Hz, 1H), 3.43 (B of AB, d, J=10.4 Hz,1H), 3.35 (s, 3H), 2.91 (s, 3H), 2.72 (A of AB, d, J=13.9 Hz, 1H), 2.66((B of AB, d, J=13.9 Hz, 1H), 1.57 (d, J=7.0 Hz, 3H). HRMS calculatedfor C₂₈H₃₄FN₃O₅S+H: 544.2276, found: 544.2275

Example 23-{[2-amino-3-hydroxy-2-(thien-3-ylmethyl)propoxy]methyl}-N-[(1R)-1-(4-fluorophenyl)ethyl]-5-[methyl(methylsulfonyl)amino]benzamide

To a solution of intermediate A3-(bromomethyl)-N-[(1R)-1-(4-fluorophenyl)ethyl]-5-[methyl(methylsulfonyl)amino]benzamide(50 mg, 0.11 mmol) in 1 mL CH₂Cl₂ cooled to 0° C. was added intermediatem tert-butyl2-hydroxy-1-(hydroxymethyl)-1-(thien-3-ylmethyl)ethylcarbamate (81 mg,0.28 mmol), silver triflate (72 mg, 0.28 mmol) and2,6-ditert-butylpyridine (76 μL, 0.34 mmol). The reaction mixture wasstirred at 0° C. for 40 min and purified by flash chromatography (20 gsilica, 30→80% EtOAc/hexanes) and by preparative HPLC (5% to 95% CH₃CNin water containing 0.1% TFA, C18 PRO YMC 20×150 mm) to affordtert-butyl2-({3-({[(1R)-1-(4-fluorophenyl)ethyl]amino}carbonyl)-5-[methyl(methylsulfonyl)amino]benzyl}oxy)-1-(hydroxymethyl)-1-(thien-3-ylmethyl)contaminated with 2,6-ditert-butylpyridine. Treatment with 10% TFA inCH₂Cl₂ for 30 min, followed by concentration under a stream of nitrogen,and purification by preparative HPLC (5% to 95% CH₃CN in watercontaining 0.1% TFA, C18 PRO YMC 20×150 mm) afforded3-{[2-amino-3-hydroxy-2-(thien-3-ylmethyl)propoxy]methyl}-N-[(1R)-1-(4-fluorophenyl)ethyl]-5-[methyl(methylsulfonyl)amino]benzamideas a TFA salt. ¹H NMR (400 MHz, d₄-MeOH) δ 8.86 (d, J=7.9 Hz, 1H), 7.83(s, 1H), 7.80 (s, 1H), 7.65 (s, 1H), 7.48-7.36 (m, 3H), 7.17 (s, 1H),7.10-7.00 (m, 2H), 6.95 (d, J=5.0 Hz, 1H), 5.30-5.20 (m, 1H), 4.68 (A ofAB, d, J=12.0 Hz, 1H), 4.63 (B of AB, d, J=12.0 Hz, 1H), 3.57 (s, 2H),3.51 (A of AB, d, J=11.0 Hz, 1H), 3.45 (B of AB, d, J=11.0 Hz, 1H), 3.09(s, 3H), 3.04 (A of AB, d, J=13.6 Hz, 1H), 3.45 (B of AB, d, J=13.6 Hz,1H), 2.94 (s, 3H), 1.58 (d, J=7.1 Hz, 3H).

HRMS calculated for C₂₆H₃₂FN₃O₅S₂+H: 550.1840, found: 550.1814.

Example 33-{[(2-amino-2-benzyl-3-hydroxypropyl)amino]methyl}-N-[(1R)-1-(4-fluorophenyl)ethyl]-5-[methyl(methylsulfonyl)amino]benzamide

Step A: Alkylation of Intermediate IV with Bromide Intermediate A.

A solution of bromide intermediate A (102 mg, 0.23 mmol), amineintermediate IV (72 mg, 0.23 mmol) and potassium carbonate (35 mg, 0.25mmol) in 1.5 mL DMF was stirred at room temperature for 16 h. Thereaction mixture was diluted with EtOAc, washed with water (2×), brine,dried over sodium sulfate and concentrated in vacuo to provide crudemethylN-(tert-butoxycarbonyl)-alpha-[({3-{[(1R)-1-(4-fluorophenyl)ethyl]amino}carbonyl)-5-[methyl(methylsulfonyl)amino]benzyl}amino)methyl]phenylalaninatewhich was used as is in step B.

Step B: Reduction of Methyl Ester

To a solution of crude methyl ester from step B (38 mg, 0.06 mmol) in 1mL THF cooled to 0° C. was added LiBH₄ (0.14 mL, 0.29 mmol, 2 M THP)dropwise. The reaction mixture was allowed to warm to room temperatureand progress was monitored by LC/MS. Additional portions of LiBH₄ wereadded over a period of 48 h to obtain disappearance of starting ester.The reaction mixture was quenched with acetone and MeOH and stirred atroom temperature for 16 h. Concentration in vacuo and purification byflash chromatography (20 g silica, 50→100% EtOAc/hexanes, followed by0→8% (10% NH₄OH in MeOH)/CH₂Cl₂) and by preparative HPLC (5% to 95%CH₃CN in water containing 0.1% TFA, C18 PRO YMC 20×150 mm) affordedtert-butyl1-benzyl-2-({3-({[(1R)-1-(4-fluorophenyl)ethyl]amino}carbonyl)-5-[methyl(methylsulfonyl)amino]benzyl}amino)-1-(hydroxymethyl)ethylcarbamatewhich was deprotected as is in step C.

Step C: Boc Removal

Treatment of Boc derivative from step B with 50% TFA in CHCl₃ for 30min, followed by concentration under a stream of nitrogen, andpurification by preparative HPLC (5% to 95% CH₃CN in water containing0.1% TFA, C18 PRO YMC 20×150 mm) afforded3-{[(2-amino-2-benzyl-3-hydroxypropyl)amino]methyl}-N-[(1R)-1-(4-fluorophenyl)ethyl]-5-[methyl(methylsulfonyl)amino]benzamideas a bis TFA salt. ¹H NMR (400 MHz, d₄-MeOH) δ 8.85 (d, J=7.9 Hz, 1H),7.84 (s, 2H), 7.67 (s, 1H), 7.46-7.38 (m, 2H), 7.36-7.25 (m, 3H),7.25-7.18 (m, 2H), 7.10-7.00 (m, 2H), 5.40-5.18 (m, 1H), 4.12-4.00 (m,2H), 3.66 (A of AB, d, J=11.0 Hz, 1H), 3.61 (B of AB, d, J=11.0 Hz, 1H),3.35 (s, 3H), 3.06-2.88 (m, 2H), 2.95 (s, 3H), 1.58 (d, J=7.1 Hz, 3H).

Example 43′-[(2-amino-2-benzyl-3-hydroxypropoxy)methyl]-5′-{[2-(2-furyl)pyrrolidin-1-yl]carbonyl}-1,1′-biphenyl-2-carbonitrile

Step A: Alkylation of Intermediate II with Bromide Intermediate B.

Alkylation oftert-butyl[1-benzyl-2-hydroxy-1-(hydroxymethyl)ethyl]carbamate II withmethyl 5-(bromomethyl)-2′-cyano-1,1′-biphenyl-3-carboxylate B usingsilver triflate and polymer-bound 2,6-ditert-butylpyridine polymer-boundfollowing a similar procedure as described in example 2 provided methyl5-({2-benzyl-2-[(tert-butoxycarbonyl)amino]-3-hydroxypropoxy}methyl)-2′-cyano-1,1′-biphenyl-3-carboxylatecontaminated with product resulting from additional transesterificationof the diol on the methyl ester. The mixture was used as is in step B.

Step B: Hydrolysis

To a solution of crude methyl5-({2-benzyl-2-[(tert-butoxycarbonyl)amino]-3-hydroxypropoxy}methyl)-2′-cyano-1,1′-biphenyl-3-carboxylatefrom step A (442 mg, 0.83 mmol) in 4 mL THP was added 1 N LiOH (1.25 mL,1.25 mmol) and the reaction mixture was stirred at room temperature for1.5 h. 1 N LiOH (1.25 mL, 1.25 mmol) was added and the reaction mixturewas stirred at room temperature for 16 h. The reaction mixture wasdiluted with water, extracted with CH₂Cl₂ (3×), dried over sodiumsulfate, concentrated in vacuo and purified by flash chromatography (90g silica, 0→85% (0.5% HOAc in EtOAc)/hexanes) to provide 259 mg of5-({2-benzyl-2-[(tert-butoxycarbonyl)amino]-3-hydroxypropoxy}methyl)-2′-cyano-1,1′-biphenyl-3-carboxylicacid as a colorless oil. ¹H NMR (400 MHz, CDCl₃) δ 8.21 (s, 1H), 8.13(s, 1H), 7.84 (s, 1H), 7.81 (d, J=7.7 Hz, 1H), 7.70 (t, J=7.3 Hz, 1H),7.57 (d, J=7.7 Hz, 1H), 7.51 (t, J=7.3 Hz, 1H), 7.30-7.16 (m, 5H), 4.98(s, 1H), 4.69 (A of AB, d, J=12.0 Hz, 1H), 4.65 (B of AB, d, J=12.0 Hz,1H), 3.77 (A of AB, d, J=12.0 Hz, 1H), 3.69 (B of AB, d, J=12.0 Hz, 1H),3.59 (A of AB, d, J=9.3 Hz, 1H), 3.69 (B of AB, d, J=9.3 Hz, 1H), 3.17(A of AB, d, J=13.8 Hz, 1H), 2.93 (B of AB, d, J=13.8 Hz, 1H), 1.44 (s,9H).

Step C: Coupling of P₃ Amine and Deprotection.

To a solution of acid from step B (20 mg, 0.04 mmol),2-(2-furyl)pyrrolidine (16 mg, 0.12 mmol, WO 2000058283) anddiisopropylethyl amine (0.017 mL, 0.1 mmol) in 0.5 DL DMF was added BOPreagent (0.021 mg, 0.05 mmol) and the reaction mixture was kept at roomtemperature for 0.25 h. Purification by preparative HPLC (5% to 95%CH₃CN in water containing 0.1% TFA, C18 PRO YMC 20×150 mm) affordedtert-butyl1-benzyl-2-[(2′-cyano-5-{[2-(2-furyl)pyrrolidin-1-yl]carbonyl}-1,1′-biphenyl-3-yl)methoxy]-1-(hydroxymethyl)ethylcarbamateas a TFA salt. Treatment with HCl(g) in CH₂Cl₂ for 16 h, followed byconcentration under a stream of nitrogen, and purification bypreparative HPLC (5% to 95% CH₃CN in water containing 0.1% TFA, C18 PROYMC 20×150 mm) and by flash chromatography (8 g silica, 0→20% (10% NH₄OHin MeOH)/CH₂Cl₂) afforded3′-[(2-amino-2-benzyl-3-hydroxypropoxy)methyl]-5′-{[2-(2-furyl)pyrrolidin-1-yl]carbonyl}-1,1′-biphenyl-2-carbonitrile.¹H NMR (400 MHz, d₄-MeOH) ca. 1:1 rotomers mixture δ 9.91-7.12 (m, 13H),6.34 (br s, 0.5; H), 6.31 (br s, 0.5H), 6.14 (br s, 0.5; H), 5.82 (br s,0.5; H), 5.42-5.36 (m, 0.5; H), 5.13-5.05 (m, 0.5; H), 4.67 (s, 1H),4.59 (A of AB, d, J=12.8 Hz, 0.5H), 4.53 (B of AB, d, J=12.8 Hz, 0.5H),3.85-3.73 (m, 1.5H), 3.65-3.54 (m, 0.5H), 3.50-3.40 (m, 2H), 2.80 (A ofAB, d, J=13.2 Hz, 1H), 2.74 (B of AB, d, J=13.2 Hz, 1H), 2.38-2.00 (m,4H). LC/MS M+H=536.

The following compounds are prepared in a manner similar to thecompounds of the foregoing examples using appropriate starting materialsand reagents.

While the invention has been described and illustrated with reference tocertain particular embodiments thereof, those skilled in the art willappreciate that various adaptations, changes, modifications,substitutions, deletions, or additions of procedures and protocols maybe made without departing from the spirit and scope of the invention. Itis intended, therefore, that the invention be defined by the scope ofthe claims that follow and that such claims be interpreted as broadly asis reasonable.

1. A compound of formula (I):

wherein: X is O or NH; Y is CH; R¹ is aryl selected from the groupconsisting of phenyl and napthyl, wherein said aryl is unsubstituted orsubstituted with one or more (a) halo, (c) —C₂₋₆ alkenyl, (d) —C₂₋₆alkynyl, (e) —OH, (f) —CN, or R² is selected from the group consistingof: (1) R⁴—S(O)₂N(R⁷)—, wherein R⁴ is C₁₋₆alkyl, wherein said alkyl isunsubstituted or substituted with one or more (a) halo, (b) —C₁₋₆alkyl,(c) —OH, (d) —CN, or (e) —O—C₁₋₆alkyl; and R⁷ is selected from the groupconsisting of (a) hydrogen, and (b) —C₁₋₆alkyl, wherein said alkyl isunsubstituted or substituted with one or more (i) halo, (ii) —C₁₋₆alkyl,(iii) OH, (iv) —CN, or (v) —O—C₁₋₆alkyl;

R³ is selected from the group consisting of:

wherein R⁵ is C₁₋₆alkyl, C₂₋₆ alkenyl or C₂₋₆ alkynyl; R^(6a), R^(6b),and R^(6c) are independently selected from the group consisting of: (1)hydrogen, (2) halo, (3) —C₁₋₆alkyl, (4) —C₂₋₆ alkenyl, (5) —C₂₋₆alkynyl, (6) —OH, (7) —CN, and (8) —O—C₁₋₆ alkyl; R⁹ and R¹⁰ areindependently selected from the group consisting of: (1) hydrogen, and(2) C₁₋₆alkyl, (3) —C₂₋₆ alkenyl, and (4) —C₂₋₆ alkynyl, or R⁹ and R¹⁰are joined together with the nitrogen atom to which they are attached toform a pyrrolidine ring, which is optionally substituted with (a) C₁₋₆alkyl, (b) —C₂₋₆ alkenyl, (c) —C₂₋₆ alkynyl, (d) (CH₂)_(n)-phenyl, and(e) (CH₂)_(n)-furanyl; wherein said alkyl, phenyl and furanyl areunsubstituted or substituted with one or more i) halo, ii) —C₁₋₆ alkyl,iii) —OH, iv) —CN, or v) —O—C₁₋₆ alkyl; and R¹¹ is selected from thegroup consisting of (1) —CH—, (2) —O—, and (3) NH—, provided that whenR¹¹ is —CH— the dotted line forms a bond and when R¹¹ is —O— or —NH— thedotted line is absent; R¹² is hydrogen, C₁₋₆ alkyl, C₂₋₆ alkenyl or C₂₋₆alkynyl; m is 1 or 2; n is 0, 1, 2, 3 or 4; p is 1, 2, 3 or 4; andpharmaceutically acceptable salts thereof.
 2. The compound of claim 1,wherein m is 1 and R¹ is phenyl unsubstituted or substituted with one ormore chloro or fluoro.
 3. The compound of claim 1, wherein m is 2 and R¹is phenyl unsubstituted or substituted with one or more chloro orfluoro.
 4. The compound of claim 1, wherein R² is (R⁴)—S(O)₂N(R⁷)— andR⁷ is C₁₋₆ alkyl.
 5. The compound of claim 4 wherein R⁴ and R⁷ are eachmethyl.
 6. The compound of claim 1, wherein R² is


7. The compound of claim 1 wherein R³ is


8. The compound of claim 7 wherein R⁵ is methyl.
 9. The compound ofclaim 1 wherein R³ is

and R⁹ and R¹⁰ are joined together with the nitrogen atom to which theyare attached to form a pyrrolidine ring which is unsubstituted orsubstituted with (a) C₁₋₆alkyl, (b) (CH₂)_(n)-phenyl, or (c)(CH₂)_(n)-furanyl.
 10. The compound of claim 9 wherein R⁹ and R¹⁰ arejoined together with the nitrogen atom to which they are attached toform a pyrrolidine ring which is substituted with —(CH₂)_(n)-furanylwherein n is
 0. 11. The compound of claim 10, wherein R³ is


12. The compound of claim 1 wherein R³ is


13. The compound of claim 1 of formula II:

wherein x, Y, R¹, R², R⁵, R^(6a), R^(6b), R^(6c) and mare as defined inclaim
 1. 14. The compound of claim 1 of formula (III):

wherein X, Y, R¹, R², R⁹, R¹⁰ and m are as defined in claim
 1. 15. Thecompound of claim 1 of formula (IV):

wherein X, Y, R¹, R², R¹¹, R¹² and mare as defined in claim
 1. 16. Thecompound of claim 1 which is selected from the group consisting of:

and pharmaceutically acceptable salts thereof.
 17. A pharmaceuticalcomposition comprising an effective amount of a compound of claim 1 anda pharmaceutically acceptable carrier.